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A DNA break– and phosphorylation-dependent positive feedback loop promotes immunoglobulin class-switch recombination

Nature Immunology volume 14, pages 1183–1189 (2013)Cite this article

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Abstract

The ability of activation-induced cytidine deaminase (AID) to efficiently mediate class-switch recombination (CSR) is dependent on its phosphorylation at Ser38; however, the trigger that induces AID phosphorylation and the mechanism by which phosphorylated AID drives CSR have not been elucidated. Here we found that phosphorylation of AID at Ser38 was induced by DNA breaks. Conversely, in the absence of AID phosphorylation, DNA breaks were not efficiently generated at switch (S) regions in the immunoglobulin heavy-chain locus (Igh), consistent with a failure of AID to interact with the endonuclease APE1. Additionally, deficiency in the DNA-damage sensor ATM impaired the phosphorylation of AID at Ser38 and the interaction of AID with APE1. Our results identify a positive feedback loop for the amplification of DNA breaks at S regions through the phosphorylation- and ATM-dependent interaction of AID with APE1.

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Figure 1: Catalytically inactive AID bound to S regions is not efficiently phosphorylated.
Figure 2: Phosphorylation of AID at Ser38 is induced by DNA breaks.
Figure 3: AicdaS38A/S38A B cells have fewer DSBs.
Figure 4: AicdaS38A/S38A B cells have less colocalization of Igh with γ-H2AX foci.
Figure 5: AID interacts with APE1.
Figure 6: ATM is required for the phosphorylation of AID bound to S regions and the interaction of AID with APE1.

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Acknowledgements

We thank J. Hurwitz (Memorial Sloan-Kettering Cancer Center) for recombinant human RPA; members of the Chaudhuri laboratory, D. Sant' Angelo and L. Denzin for critically reviewing the manuscript and for discussions and suggestions; V. Bermudez and J. Hurwitz for help in purifying recombinant human RPA; J. Lange and S. Keeney (Memorial Sloan-Kettering Cancer Center) for Atm−/− mice; and D.M. Wilson III (US National Institutes of Health) for the plasmid for expression of human APE1. Supported by the US National Institutes of Health (RO1AI23283 and R21 AI099908 to J.S., 5 R01 CA138646-04 to K.D.M. and 1RO1AI072194 to J.C.) and the Starr Cancer Consortium (I4-A447 to J.C.).

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  1. Kayleigh Herrick-Reynolds, Bharat Vaidyanathan and Joseph N Pucella: These authors contributed equally to this work.

Authors and Affiliations

  1. Immunology Program, Memorial Sloan-Kettering Cancer Center, Gerstner Sloan-Kettering Graduate School, New York, New York, USA

    Bao Q Vuong, Kayleigh Herrick-Reynolds, Bharat Vaidyanathan, Joseph N Pucella, Laura Nicolas, Urszula Nowak, Numa Rahman & Jayanta Chaudhuri

  2. Immunology and Microbial Pathogenesis Program, Weill-Cornell Graduate School of Medical Sciences, New York, New York, USA

    Bharat Vaidyanathan, Urszula Nowak & Jayanta Chaudhuri

  3. Department of Microbiology & Physiological Systems, University of Massachusetts Medical School, Worcester, Massachusetts, USA

    Anna J Ucher & Janet Stavnezer

  4. The Jackson Laboratory, Bar Harbor, Maine, USA

    Nina M Donghia & Kevin D Mills

  5. Yale Cancer Center, Yale University School of Medicine, New Haven, Connecticut, USA

    Xiwen Gu & Matthew P Strout

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Contributions

B.Q.V. and J.C. conceived of the study and wrote the paper; B.Q.V., K.H.-R., B.V., J.N.P. and U.N. did the ChIP experiments; B.Q.V., K.H.-R. and B.V. did the coimmunoprecipitation and in vitro binding experiments; A.J.U. and J.S. did the LM-PCR experiment and bred the Apex1+/−Apex2−/− mice; N.M.D. and K.D.M. did the immuno-FISH; X.G. and M.P.S. bred the Ung−/−Msh2−/− mice; and N.R., B.Q.V., K.H.-R. and L.N. bred and analyzed the Aicda−/−, Atm−/− and AicdaS38A/S38A mice.

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Correspondence to Janet Stavnezer or Jayanta Chaudhuri.

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Vuong, B., Herrick-Reynolds, K., Vaidyanathan, B. et al. A DNA break– and phosphorylation-dependent positive feedback loop promotes immunoglobulin class-switch recombination. Nat Immunol 14, 1183–1189 (2013). https://doi.org/10.1038/ni.2732

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